MUC1 is a transmembrane mucin that is highly expressed in various cancers and correlates with malignant potential. Important cancer-related genes such as p16 and E-cadherin are controlled epigenetically; however, MUC1 has been overlooked in epigenetics. Herein, we provide the first report that MUC1 gene expression is regulated by DNA methylation and histone H3 lysine 9 (H3-K9) modification of the MUC1 promoter. The recently developed MassARRAY assay was performed to investigate the DNA methylation status of 184 CpG sites from À2,753 to +263. Near the transcriptional start site, the DNA methylation level of MUC1-negative cancer cell lines (e.g., MDA-MB-453) was high, whereas that of MUC1-positive cell lines (e.g., MCF-7) was low. Histone H3-K9 modification status was also closely related to MUC1 gene expression. Furthermore, MUC1 mRNA expression in MUC1-negative cells was restored by treatment with the DNA methylation inhibitor 5-aza-2 ¶-deoxycytidine. Our results indicate that DNA methylation and histone H3-K9 modification in the 5 ¶ flanking region play a critical role in MUC1 gene expression, and this study defines MUC1 as a new member of the class of epigenetically controlled genes. An understanding of the epigenetic changes of MUC1 may be of importance for diagnosis of carcinogenic risk and prediction of outcome for cancer patients.

Mucin 4 (MUC4) is a high molecular weight transmembrane mucin that is overexpressed in many carcinomas and is a risk factor associated with a poor prognosis. In this study, we show that the DNA methylation pattern is intimately correlated with MUC4 expression in breast, lung, pancreas and colon cancer cell lines. We mapped the DNA methylation status of 94 CpG sites from À3622 to þ 29 using MassARRAY analysis that utilises base-specific cleavage of nucleic acids. MUC4-negative cancer cell lines and those with low MUC4 expression (eg, A427) were highly methylated near the transcriptional start site, whereas MUC4-positive cell lines (eg, NCI-H292) had low methylation levels. Moreover, 5-aza-2 0 -deoxycytidine and trichostatin A treatment of MUC4-negative cells or those with low MUC4 expression caused elevation of MUC4 mRNA. Our results suggest that DNA methylation in the 5 0 flanking region play an important role in MUC4 gene expression in carcinomas of various organs. An understanding of epigenetic changes in MUC4 may contribute to the diagnosis of carcinogenic risk and prediction of outcome in patients with cancer.

In addition to conventional cytology, liquid-based cytology (LBC) is also used for immunocytochemistry and gene analysis. However, an appropriate method to obtain high quality DNA for next-generation sequencing (NGS) using LBC specimens remains controversial. We determined the optimal conditions for fixation with an alcohol-based fixative for LBC and DNA extraction using cultured cancer cell lines and clinical specimens. The extracted DNA was processed for NGS after the DNA quality was confirmed based on the DNA concentration and degree of degradation. The optimal conditions for cultured cells to obtain high quality DNA were to fix the cells at a density of 6 × 10 3 or 2 × 10 4 cells/mL and to use the magnetic bead-based DNA extraction method. Even after storing the fixed cells for 90 days, DNA extracted using the above and other extraction kits, including membrane-based methods, did not undergo degradation. Furthermore, 5-year-old residual LBC samples demonstrated high DNA quality that was suitable for NGS. Furthermore, a cancer genome panel analysis was successfully performed with DNA extracted from cultured cells fixed at 6 × 10 3 cells/mL for 90 days, and with DNA from residual LBC samples even after 1 year of storage. Residual LBC samples may be a useful source of DNA for clinical NGS to promote genome-based cancer medicine.

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Our results indicate that MUC5AC is regulated by CpG methylation and histone H3-K9 modification of the MUC5AC promoter distal region, but not by miRNAs. An understanding of the epigenetic regulation of MUC5AC may be of importance for the diagnosis of carcinogenic risk in the pancreatobiliary system.

We present distance measurement of the semiregular variable RX Bootis (RX Boo) with its annual parallax. Using the unique dual-beam system of the VLBI Exploration of Radio Astrometry (VERA) telescope, we conducted astrometric observations of a water maser spot accompanying RX Boo referred to the quasar J1419+2706 separated by 1 • .69 from RX Boo. We have measured the annual parallax of RX Boo to be 7.31 ± 0.50 mas, corresponding to a distance of 136 +10 −9 pc, from the one-year monitoring observation data of one maser spot at V LSR = 3.2 km s −1 . The distance itself is consistent with the one obtained with Hipparcos. The distance uncertainty is reduced by a factor of two, allowing us to determine the stellar properties more accurately. Using our distance, we discuss the location of RX Boo in various sequences of Period-Luminosity (PL) relations. We found RX Boo is located in the Mira sequence of PL relation. In addition, we calculated the radius of photosphere and the mass limitation of RX Boo and discussed its evolutionary status.

Abstract-Effect of etizolam on platelet activating factor (PAF) and N-formyl-

We use astrometry to measure the distances to Galactic Mira variable stars. Our purpose is to determine a precise period-luminosity relation (PLR). At present, we do not have a precise PLR for Galactic Mira stars because of the large uncertainties affecting their distance estimates. To reduce the uncertainties, we adopted the Very Long Baseline Interferometry method and measured annual parallaxes of Mira variable stars with VERA. In addition to our previous results, we obtained three new distances for Mira variable stars. Based on our observations, the typical uncertainty in a given distance measurement is reduced to below 10%. At this conference, we present the current status of our project. To establish a precise Galactic PLR, we continue to observe more Mira variables. In addition, the apparent magnitudes of the target stars should be studied carefully.